Simple and quick determination of analgesics and other contaminants of emerging concern in environmental waters by on-line solid phase extraction coupled to liquid chromatography-tandem mass spectrometry.

A simple and quick analytical method has been developed for the determination of pharmaceutical compounds in water. An on-line solid-phase extraction (SPE) coupled to liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been optimized to determine 7 contaminants of emerging concern in environmental waters at ngL(-1) levels. This procedure requires minimal sample handling and small sample volume (900µL) with a total running time of 18min. Several SPE parameters were evaluated and optimized in order to achieve a high sample throughput. Therefore sample volume, carryover and reusability of the cartridges were evaluated. Performance characteristics were evaluated and good linearity was obtained (R(2)>0.98). Recoveries were evaluated in spiked samples at three concentrations and the values ranged from 71 to 104%. Intra and inter-day precision was lower than 10 and 13% respectively. Limits of quantification were equal to or lower than 10ngL(-1), except for 1,7-dimethylxanthine (20ngL(-1)) and ibuprofen (50ngL(-1)). The method was applied to 20 environmental water samples, and ibuprofen was the compound most widely detected at concentrations up to 42.06µgL(-1), whereas the other compounds were detected in fewer samples at lower concentrations (up to 15.99µgL(-1)).

Fast analysis of 4-tertoctylphenol, pentachlorophenol and 4-nonylphenol in river sediments by QuEChERS extraction procedure combined with GC-QqQ-MS/MS.

A quick, easy, cheap, effective, rugged and safe (QuEChERS)-based extraction method has been optimized for the determination of pentachlorophenol, 4-tertoctylphenol and 4-nonylphenol in river sediments. The extraction method was followed by gas chromatography-triple quadrupole tandem mass spectrometry (GC-QqQ-MS/MS) analysis, which ensures the reliable identification of the target compounds. The proposed method has been validated allowing the successful determination of the selected compounds, with recoveries ranging from 72 to 96%, when three concentration levels were evaluated (10, 50 and 100µgkg(-1)) and inter-day and intra-day precision, expressed as relative standard deviation (RSD), were lower than 20%. The method showed limits of detection (LODs) and limits of quantification (LOQs) ranging from 0.1 to 2.0µgkg(-1) and from 0.5 to 5.0µgkg(-1), respectively. Finally, 25 real samples from Poland have been analyzed, and only 4-tertoctylphenol was detected at concentrations up to 8.9µgkg(-1) of soil dry weight.

Food contaminant analysis at high resolution mass spectrometry: application for the determination of veterinary drugs in milk.

Veterinary drugs (VDs) can remain in milk as a consequence of their use in livestock. In order to control the levels of VD residues in milk, screening methodologies can be applied for a rapid discrimination among negative and non-negative samples. In a second stage, non-negative samples are classified as negative or positive samples by using a confirmation method. Pre-target screening methods in low resolution MS (LRMS) are normally applied, but the number of analytes is limited, whereas the information obtained by full scan acquisition in high resolution mass spectrometry (HRMS) is improved. Here, three screening methods (running time<4 min) based on Orbitrap, quadrupole-time of flight (QqTOF) and triple quadrupole (QqQ) have been compared, using in all cases ultra-high performance liquid chromatography (UHPLC). For HRMS, the identification of the VDs was based on retention time (RT) and accurate mass measurements. Confirmation was based on the monitoring of fragments generated without precursor selection. The performance characteristics of the screening method provided reliable information regarding the presence or absence of the compounds below an established value, including uncertainty region and cut-off values. Better results in terms of cut-off values (≤ 5.0 µg kg(-1), except for spiramycin with a cut-off of 13.4 µg kg(-1) for milk samples and 43.1 µg kg(-1) for powdered milk based, emamectin with a cut-off of 42.2 µg kg(-1) for milk samples and doxycycline, with a cut-off value of 15.8 µg kg(-1) in powdered milk-based infant formulae) and uncertainty region were obtained using the Orbitrap-based screening method, which was submitted to further validation and used to analyze different real milk samples. The proposed method can be used in routine analysis, providing reliable results.

Comparison of the efficiency of different extraction methods for the simultaneous determination of mycotoxins and pesticides in milk samples by ultra high-performance liquid chromatography-tandem mass spectrometry.

A rapid multi-analyte method has been developed for the simultaneous determination of pesticides and mycotoxins in milk by ultra high-performance liquid chromatography coupled to triple quadrupole mass spectrometry (UHPLC-QqQ-MS/MS). A variety of methodologies has been evaluated, including solid-phase extraction (SPE), "dilute-and-shoot" (liquid-liquid extraction-based procedures), and QuEChERS (quick, easy, cheap, effective, rugged, and safe)-based methods. The optimization and development process was carried out considering that the maximum residue level for aflatoxin M1 (AFM1) in milk in the European Union (EU) is set at 0.05 µg kg(-1), which is the lowest tolerance in the target compounds. The selected method consisted of an extraction by SPE using C18 as sorbent and methanol as elution solvent. The final determination was performed by UHPLC-QqQ-MS/MS. Matrix-matched standard calibration was used for quantification, obtaining recoveries in the range 60-120% with relative standard deviations <25%, at three spiking levels: 0.5, 10, and 50 µg kg(-1) (ten times lower for AFM1). Limits of quantification ranged from 0.20 to 0.67 µg kg(-1), which were always below or equal to the established tolerance levels by the EU. Finally, the selected method was applied to different types of milk.

Determination of ochratoxin A and T-2 toxin in alcoholic beverages by hollow fiber liquid phase microextraction and ultra high-pressure liquid chromatography coupled to tandem mass spectrometry.

A new method for the determination of ochratoxin A and T-2 toxin in alcoholic beverages (wine and beer) by hollow fiber liquid microextraction was optimized. The extraction step was followed by ultra high-pressure liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). The extraction procedure was based on the extraction of mycotoxins from the sample to the organic solvent (1-octanol) immobilized in the fiber, and afterwards, they were desorbed in a mixture of acetonitrile/water (80:20, v/v) at pH 7 prior to chromatographic determination. Different variables affecting the extraction process such as organic solvent, salt content, extraction time and desorption solution were studied. The developed method was validated in wine and beer, using white wine and alcoholic beer as representative matrices for both types of samples. Relative recoveries higher than 70% were obtained for the selected mycotoxins. Good linearity (R(2)>0.993) was obtained and quantification limits (0.02-0.09 microg L(-1)) below European regulatory levels were achieved. Repeatability, expressed as relative standard deviation, was always lower than 12%, whereas interday precision was lower than 21%. The proposed method was applied to the analysis of several types of wines and beers and ochratoxin A was detected in a rosé wine at 1.1 microg L(-1).

Comparison of ultrasonic and pressurized liquid extraction for the analysis of polycyclic aromatic compounds in soil samples by gas chromatography coupled to tandem mass spectrometry.

A pressurized liquid extraction (PLE) method has been optimized for the determination of polycyclic aromatic hydrocarbons (PAHs) in soil samples and it was compared with ultrasonic extraction. The extraction step was followed by gas chromatography-triple quadrupole mass spectrometry (GC-QqQ-MS/MS) analysis. Parameters such as type of solvent, extraction time, extraction temperature and number of extractions were optimized. There were no significant differences among the two extraction methods although better extraction efficiencies were obtained when PLE was used, minimizing extraction time and solvent consumption. PLE procedure was validated, obtaining limits of detection (LODs) ranging from 0.02 to 0.75 microg kg(-1) and limits of quantification (LOQs) ranging from 0.07 to 2.50 microg kg(-1) for the selected PAHs. Recoveries were in the range of 59-110%, except for naphthalene, which was the most volatile PAH. Finally, the method was applied to real soil samples from Southeast of Spain. PAHs concentrations were low, and phenanthrene, pyrene, fluorene, benzo[a]pyrene and chrysene were the most frequently detected analytes in the samples.

Application of hollow fibre liquid phase microextraction for the multiresidue determination of pesticides in alcoholic beverages by ultra-high pressure liquid chromatography coupled to tandem mass spectrometry.

An alternative method has been developed to determine more than 50 pesticides in alcoholic beverages using hollow fibre liquid phase microextraction (HF-LPME) followed by ultra-high pressure liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS), without any further clean-up step. Pesticides were extracted from the sample to the organic solvent immobilized in the fibre and they were desorbed in methanol prior to chromatographic analysis. Experimental parameters related to microextraction such as type of organic solvent, extraction time and agitation rate have been optimized. The extraction method has been validated for several types of alcoholic beverages such as wine and beer, and no matrix effect was observed. The technique requires minimal sample handling and solvent consumption. Using optimum conditions, low detection limits (0.01-5.61microgL(-1)) and good linearity (R(2)>0.95) were obtained. Repeatability and interday precision ranged from 3.0 to 16.8% and from 5.9 to 21.2%, respectively. Finally the optimized method was applied to real samples and carbaryl, triadimenol, spyroxamine, epoxiconazole, triflumizol and fenazaquin were detected in some of the analyzed samples. The obtained results indicated that the new method can be successfully applied for extraction and determination of pesticides in alcoholic beverages, increasing sample throughput.

Application of gas chromatography-triple quadrupole mass spectrometry in the quantification-confirmation of pesticides and polychlorinated biphenyls in eggs at trace levels.

A new multiresidue method has been developed and validated for the simultaneous analysis of 57 compounds, including organochlorine and organophosphorus pesticide residues (OCPs and OPPs) and polychlorinated biphenyls (PCBs), in eggs at trace levels by gas chromatography coupled to triple quadrupole mass spectrometry (GC-QqQ-MS/MS). Egg samples were extracted by a simple and fast matrix solid phase dispersion (MSPD) procedure using C18 as sorbent, and ethyl acetate and acetonitrile saturated in n-hexane (85:15, v/v) as elution solvent with a simultaneous clean up with Florisil in-line. The QqQ analyzer acquired data in selected reaction monitoring (SRM) mode, permitting both quantification and confirmation in a single injection with a running time reduced up to 17.70 min. Recovery was in the range of 70-110% and 70-106% at 15 and 50 microg/kg, respectively. Precision values expressed as relative standard deviation (RSD) were lower than 20%. Linearity in the range of 10-150 microg/kg provided determination coefficients (R(2)) higher than 0.98 for all compounds. Limits of detection (LODs) for pesticides were < or =2.25 microg/kg and limits of quantification (LOQs) ranged from 0.02 to 7.78 microg/kg. LODs for PCBs were < or =0.41 microg/kg and LOQ were < or =0.71 microg/kg. The method was applied to real samples. Endosulfan sulphate and p,p'-DDE were found in two samples at concentrations below the first calibration level.

Assessment of butyltin and phenyltin pollution in the sea mullet, Mugil cephalus, along the Moroccan and Spanish coasts (Mediterranean Sea).

Levels of three butyltin (BTs) compounds, monobutyltin (MBT), dibutyltin (DBT) and tributyltin (TBT), and three phenyltin (PTs) compounds, monophenyltin (MPhT), diphenyltin (DPhT), and triphenyltin (TPhT), were determined in the sea mullet, Mugil cephalus, collected along the northern Mediterranean coast of Morocco, and the south Mediterranean coast of Spain. Also, sediment samples from the Guadalquivir River (Spain) near to its mouth in the Mediterranean Sea were analysed. TBT and TPhT were the predominant compounds in the analyzed samples, and TBT concentrations were higher in liver than in muscle. In general, the total content of BTs in these samples was higher than PT levels. In the Moroccan coast, the highest value of BTs (18.00 mg/Kg wet weight) was found in the west harbour of the Tangier site, while the lowest concentration (0.01 mg/Kg wet weight) was detected in the Oued Laoue site. Concerning PTs compounds, the highest value (4.00 mg/Kg wet weight) was found in the harbour of the M'diq Site and the lowest value (0.01 mg/Kg wet weight) was detected in the Oued Laoue site. On the other hand, organotin (OT) residues were not detected in the water samples of the Spanish coast although TBT was identified in four of the nine sediment samples from the Guadalquivir River at concentrations between 15.45 and 245.07 mg/Kg wet weight.

Evaluation of different sample treatments for determining pesticide residues in fat vegetable matrices like avocado by low-pressure gas chromatography-tandem mass spectrometry.

A multi-residue method has been developed for determining 65 pesticide residues in greasy vegetable matrices such as avocado. Conventional organic solvent extraction assisted by a high-speed homogenizer was compared to pressurized liquid extraction (PLE) as extraction techniques. Following this, the lipophilic extract was purified using gel permeation chromatography (GPC). Alternative clean-up methods were also evaluated, as solid-phase extraction cartridges individually used and downstream coupled, but less effective lipophilic separation was archived. The pesticide residue determination was carried out using low-pressure gas chromatography coupled to tandem mass spectrometry (LP-GC-MS-MS), showing the applicability of this type of GC columns for the analysis of fat vegetable matrices. The proposed methodology was validated in avocado matrix. The recoveries were in the range 70-110%, with RSD values lower than 19%, at 12 and 50 microg/kg spiking levels. The limits of quantitation (LOQs) were in the range 0.04-8.33 microg/kg and the limits of detection (LODs) were between 0.01 and 2.50 microg/kg. All of them were lower than the maximum residue levels (MRLs) set by the European Union (EU) in avocado. The proposed method was evaluated analyzing pesticide residues in real avocado samples.

Determination of ascorbic acid and carotenoids in food commodities by liquid chromatography with mass spectrometry detection.

Two methods, one to determine ascorbic acid and one to determine lycopene and beta-carotene, in vegetables and fruits by liquid chromatography coupled with mass spectrometry (LC-MS) have been established. The chromatographic separation of the studied compounds and their MS parameters were optimized to improve selectivity and sensitivity. In both methods, separation was carried out with two coupled columns, first a C(18) and then a dC(18), using as mobile phase 70% methanol (0.005% acetic acid) and 30% acetic acid 0.05% for ascorbic acid determination and a mixture of methanol, tetrahydrofuran, and acetonitrile (60:30:10 v/v/v) for carotenoid analysis in isocratic mode. The molecular ion was selected for the quantification in selective ion monitoring (SIM) mode. Ascorbic acid was detected with electrospray ionization probe (ESI) in negative mode, while chemical ionization atmospheric pressure (APCI) in positive mode was used for the target carotenoids. The methodology for ascorbic acid analysis is based on an extraction with polytron using methanol and a mixture of methaphosphoric acid and acetic acid. Extraction of the carotenoids was carried out with tetrahydrofuran/methanol (1:1) (v/v). The proposed methods were applied, after their corresponding validations, to the analysis of four varieties of tomatoes, tomato in tin enriched and dried tomato, and to the analysis of mango and kiwi fruits, to compare the content in these compounds. Moreover, the influence of the process of freezing and the effect that the manipulation/preservation has in the content of ascorbic acid in tomato have also been studied.

Application of internal quality control to the analysis of quaternary ammonium compounds in surface and groundwater from Andalusia (Spain) by liquid chromatography with mass spectrometry.

A method has been developed for the simultaneous determination of paraquat (PQ), deiquat (DQ), chlormequat (CQ) and mepiquat (MQ) in water samples by liquid chromatography (LC) coupled with electrospray ionization mass spectrometry (MS). The LC separations of the target compounds, as well as their MS parameters, were optimized in order to improve selectivity and sensitivity. Separation was carried out in a Xterra C8 column, using as mobile phase methanol-heptafluorobutyric acid (HFBA) in isocratic mode. The molecular ion was selected for the quantitation in selected ion monitoring (SIM) mode. Off-line solid-phase extraction (SPE) was applied with silica cartridges in order to preconcentrate the compounds from waters. Detection limits were in the range 0.02-0.40 microg l(-1). Recovery range varied between 89 and 99.5% with precision values lower than 6%. The method has been applied successfully to the analysis of both surface and groundwater samples from agricultural areas of Andalusia (Spain), using well defined internal quality control (IQC) criteria. The results revealed the presence of deiquat and paraquat in some samples.

Monitoring multi-class pesticide residues in fresh fruits and vegetables by liquid chromatography with tandem mass spectrometry.

A new analytical method was developed using liquid chromatography with tandem mass spectrometry for the routine analysis of 31 multi-class pesticide residues and applied to approximately 50 fresh fruit and vegetable samples (green bean, cucumber, pepper, tomato, eggplant, watermelon, melon and zucchini). Extraction of the pesticides with ethyl acetate was carried out. The optimal ionisation conditions were selected for each pesticide in the same run. The procedure was validated and the values of some merit figures, such as recovery, precision, linear range, detection limit and quantification limit for each pesticide were calculated together with its calculated expanded uncertainty (U). The average recoveries in cucumber obtained for each pesticide ranged between 74 and 105% at two different fortification levels (n = 10 each) that ranged between 9 and 250 ng g(-1) (depending on the pesticide). The uncertainty associated to the analytical method was lower than 23% for all compounds tested. The calculated limits of detection and quantitation were typically <1 ng g(-1) that were much lower than the maximum residue levels established by European legislation.

Single step determination of fragrances in Cucurbita flowers by coupling headspace solid-phase microextraction low-pressure gas chromatography-tandem mass spectrometry.

Coupling headspace solid-phase microextraction (HS-SPME) and low-pressure gas chromatography-tandem mass spectrometry (LP-GC-MS-MS) has been used for determining 20 volatile compounds present in flowers. HS-SPME coupled with LP-GC-MS-MS acts in a synergic way allowing a fast extraction and analysis of the target compounds. The method has been optimised studying the influence of the adsorption temperature and adsorption time. The best results were obtained heating the SPME vials at 60 degrees C for 5 min using 65 microm poly(dimethylsiloxane-divinylbenzene) fibers. The validation of the method ensures the fitness for the purpose of the analytical method, achieving appropriate lower limits, recoveries and precision. The analytical method has been applied to the characterisation of zucchini flowers fragrances in air using passive sampling, in order to improve our knowledge on zucchini fragrances and to better pollination technique in future steps.

Rapid pesticide analysis, in post-harvest plants used as animal feed, by low-pressure gas chromatography-tandem mass spectrometry.

A wide range of pesticides used to control pests in vegetables have been determined in agricultural plant waste from beans, watermelons, and melons grown in greenhouses located in a predominantly agricultural area in Southeast Spain (Almería). Analysis of the pesticides was carried out by low-pressure gas chromatography (LP-GC) with mass spectrometry in tandem (MS-MS) mode, after extraction of the lyophilized samples with dichloromethane. The influence of the sample matrix on the analysis was avoided by use of matrix-matched standards. Linearity, detection limit ( LOD), quantitation limit ( LOQ), recovery, and precision for each pesticide were calculated. The most frequently encountered pesticides were endosulfan (>73% of the analyzed samples) and buprofezin (>55% of the samples), followed by cypermethrin, pirimifos-methyl, bifentrin, and chlorpyrifos (>30% of the samples). The pesticide found at the highest concentration level was endosulfan (223.33 mg kg(-1)) in a watermelon sample.

Trace determination of organotin compounds in water, sediment and mussel samples by low-pressure gas chromatography coupled to tandem mass spectrometry.

A fast method for the determination of eight organotin compounds (OTs), monobutyltin (MBT), dibutyltin (DBT), tributyltin (TBT), tetrabutyltin (TeBT), monophenyltin (MPhT), diphenyltin (DPhT), triphenyltin (TPhT) and tetraphenyltin (TePhT), in water, sediments and mussels, was developed using low-pressure gas chromatography/tandem mass spectrometry (LPGC/MS/MS). The method is based on sodium diethyldithiocarbamate (DDTC) complexation of the ionic organotins, followed by extraction of the target matrices and derivatization by a Grignard reagent, as described in a previously published method for water samples. Solid-phase extraction was selected as extraction method from water samples after comparison with liquid-liquid extraction, but extraction of the OTs from sediment and mussel samples was performed using toluene. Matrix-matched calibration standards were used to minimize matrix effects. The analytical process was validated by the analysis of spiked blank samples. Performance characteristics such as linearity, detection limit (LOD), quantitation limit (LOQ), precision, and recovery were determined. Recoveries of OTs in spiked matrices ranged from 86-108% in water and from 78-110% in sediments and mussels, with precision values lower than 18%. Detection limits ranged from 0.1-9.6 ng L(-1) in water, and 0.03-6.10 microg kg(-1) in the other matrices. The present implementation of LPGC rather than conventional capillary GC permitted use of large-volume injection and reduced analysis time by a factor of two. The proposed methodology was applied to the determination of OTs in real samples of water, marine sediments and mussels from the west coast of the Mediterranean Sea (Spain).

Application of GRAM and TLD to the resolution and quantitation of real complex multicomponent mixtures by fluorescence spectroscopy.

The application of the generalised rank annihilation method (GRAM) and the trilinear decomposition (TLD) method to the resolution and quantitation of fluorescence excitation-emission matrices of a ternary mixture of pesticides, carbendazim, fuberidazole, and thiabendazole, with overlapped spectra is described. The results obtained with both methods are compared and evaluated using measures of similarity (correlation coefficients) between the real and estimated spectra. Both approaches have been tested using augmented data matrices containing only two samples, but none of these methods succeeded completely in resolution of the system. When TLD was applied to augmented data matrices containing more than two samples better performance was achieved. To illustrate the application of both algorithms to real samples, they were used in the analysis of water samples containing the target pesticides. Again, TLD had an advantage over GRAM because the ability to analyse data from multiple (more than two) samples simultaneously allowed the resolution of the mixtures.

Determination of heavy metals in crayfish by ICP-MS with a microwave-assisted digestion treatment.

Contamination of the American red crayfish, from the Guadiamar riverside, is due to the disastrous toxic spill that occurred on 25 April 1998 in the mining area of Aznalcóllar-Los Frailes, Spain. A high concentration of heavy metals in the waters from the mine pool and their spill to the river Guadiamar was the cause of the destruction of a great number of animal and vegetable organisms. An ICP-MS method for the total determination of heavy metals (Cu, Zn, Pb, and Cd) in whole bodies of American red crayfish (Procambarus clarkii) was used. Metals were extracted from the matrix in a closed-vessel microwave digestion system with nitric acid and hydrogen peroxide. A study of the uncertainty of the method for the determination of metals was carried out; at a concentration of 5 microg/L, the uncertainty was below 34%.